Hyaluronic Acid as Capacitation Inductor: Metabolic Changes and Membrane‐Associated Adenylate Cyclase Regulation

The aim of this research was to study the effect of hyaluronic acid on bovine cryopreserved spermatozoa compared with heparin as regards the variation of capacitation induction, cellular oxidative metabolism and intracellular signal induced by membrane‐associated adenylate cyclase to propose hyaluronic acid as a capacitation inductor. Heparin or hyaluronic acid and lysophosphatidylcholine were used to induce sperm capacitation and acrosome reaction, respectively. 2′,5′‐dideoxyadenosine was used as a membrane‐associated adenylate cyclase inhibitor. The highest percentages of capacitated spermatozoa and live spermatozoa with acrosome integrity were obtained by incubating sperm for 60 min using 1000 μg/ml hyaluronic acid. In these conditions, capacitation induced by hyaluronic acid was lower compared with heparin; nonetheless both glycosaminoglycans promote intracellular changes that allow true acrosome reaction in vitro induced by lysophosphatidylcholine in bovine spermatozoa. Oxygen consumption in heparin‐capacitated spermatozoa was significantly higher than in hyaluronic acid‐treated spermatozoa. With all treatments, mitochondrial coupling was observed when a specific uncoupler of the respiratory chain was added. The inhibition of membrane‐associated adenylate cyclase significantly blocked capacitation induction produced by hyaluronic acid, maintaining a basal sperm oxygen uptake in contrast to heparin effect in which both sperm parameters were inhibited, suggesting that the membrane‐associated adenylate cyclase activation is involved in the intracellular signal mechanisms induced by both capacitation inductors, but only regulates mitochondrial oxidative phosphorylation in heparin‐capacitated spermatozoa.     

Behavioral response to selected feed attractants and stimulants in Pacific white shrimp, Litopenaeus vannamei

Nine commercial feeding attractants and stimulants for Litopenaeus vannamei were evaluated by observation of behavioral responses in animals allotted in one Y-maze aquarium apparatus. In the validation phase, fishmeal–Brazilian origin (FMBO); fishmeal–Peruvian origin (FMPO); blood meal (BM), meat and bone meal (MBM), squid meal (SM), fish oil (FO) and fish solubles (FS) were evaluated. There was also a control without stimulatory raw material. The tested materials were included at 3% in neutral gelatin pellets (wet basis). In each behavioral observation, two different ingredients were offered at the same time in equal quantities, being allotted in the end of each chamber's arm. In Phase II after system validation, the following commercial attractants were tested: 80% crude protein (CP) vegetable dried biomass (VDB₈₀), 68% CP vegetable dried biomass + glutamate + betaine (VDB₆₈), complex of amino acids (alanine, valine, glycine, proline, serine, histidine, glutamic acid, tyrosine and betaine) with enzymatically digested bivalve mollusk (CAA), condensed fish soluble protein (CFSP), squid liver meal (SLM), betaine (Bet), dried fish solubles–low biogenic amines (DFS_LB), dried fish solubles–high biogenic amines (DFS_HB) and whole squid protein hydrolysate (WSPH). Attractants were used at a 3% level wet basis with neutral gelatin, without any additional ingredient source available. The best four commercial attractants from this phase (CAA, CFSP, SLM and WSPH) were compared under 0.5% and 1.0% levels. In Phase I of the study, a higher percentage of choices were observed for FMPO and FMBO. BM and FO were the least chosen ingredients. In Phase II, the worst results were observed for Bet, DFS_HB and, mainly, for VDB₈₀ and VDB₆₈. When two-by-two comparisons were performed, results suggested that CFSP and CAA were the best commercial attractants tested. In the last phase, both CFSP and CAA at 1.0% level were significantly more chosen by shrimp than CFSP (0.5%), SLM (0.5 or 1.0%) or WSPH (0.5 or 1.0%). At both 0.5% and 1.0% levels, shrimp spent a similar amount of time feeding on CFSP and CAA. However, only CAA was statistically better than SLM and WSPH together. Further work is required to better elucidate the chemical drivers of chemostimulation for L. vannamei for each one of the attractants tested.     

Effects of the protein/carbohydrate ratio of extruded diets on protein synthesis, protein growth and body composition in juvenile brown trout (Salmo trutta)

High levels of protein in fish feeds result in higher costs and nitrogen waste. Therefore, studies focused on protein substitution by alternative energy sources are necessary. Here we examine whether the protein:carbohydrate ratio in extruded diets affects protein-turnover parameters and the main tissue components in brown trout. Juvenile fish were adapted to two extruded diets: one higher in protein and lower in carbohydrate, C: 45/28 and the other higher in carbohydrate and lower in protein, HC: 37/40. Gross energy was 19 MJ kg⁻¹ DM in both diets. Protein and lipid digestibility were high and similar in both groups (≈90%), whereas total and carbohydrate digestibility and digestible energy were lower in the HC-fed group than in the C-fed group. Consequently, when fish were adapted to the diets, plasma levels of glucose and insulin differed between diets. HC-group presented higher plasma glucose levels and lower plasma insulin levels than C-group. Protein synthesis rates in white muscle, liver and whole body did not differ significantly between diets. In contrast, protein accretion rate of white muscle and whole body were significantly lower in the HC group, indicating an increase of protein degradation in these tissues and a decrease of synthesis retention efficiency. In spite of the daily protein intake of HC group was lower than C-group, the anabolic stimulation efficiency was increased by 34% in HC group. Protein and lipid contents in white muscle and liver were stable throughout the experiment. No hepatomegalia or increased fat deposition was observed in fish fed HC. Differences in specific growth rates (C: 0.88%; HC: 0.77%) were associated more with the lower protein consumption rate and the lower level of digestible energy in fish fed HC than with the higher dietary carbohydrate content of the diet.     

Space–time patterns of co‐variation of biodiversity and primary production in phytoplankton guilds of coastal marine environments

• 1. The relevance of biodiversity to ecosystem processes is a major topic in ecology. Here, we analyse the relationship between biodiversity and productivity of the nano‐ and micro‐phytoplankton guilds in coastal marine ecosystems.
• 2. The patterns of variation of species richness, diversity and primary productivity (as ¹⁴C assimilation) were studied in two marine areas: a eutrophic–mesotrophic area beside the River Po delta (northern Adriatic) and an oligotrophic area around the Salento peninsula (southern Adriatic–Ionian). The study was carried out at 23 sites in the northern area and at 45 sites in the southern area. Sites were arranged on expected spatial and temporal gradients of primary productivity variation, according to distance from the coast, optical depths and seasonal period.
• 3. 167 taxa were identified in the northern area and 153 taxa in the southern area. In both areas, the taxonomic composition of the nano‐ and micro‐phytoplankton guilds exhibited greater temporal than spatial variation. The latter was much higher in the southern area than in the northern area (average dissimilarity between stations being 70.7±0.8% and 44.7±4.2% respectively).
• 4. Primary productivity varied in space and time on the gradients considered. Phytoplankton species richness and diversity exhibited significant patterns of variation in space and time; overall, these were inversely related to the primary productivity patterns in the northern area, whereas they were directly related in the southern area.
• 5. The small individual size and the high turnover rate of phytoplankton are likely to underlie the observed relationships, which emphasized a threshold response to nutrient enrichment in agreement with the ‘paradox of enrichment’. Under resource enrichment conditions, the high turnover of producers leads to hierarchical partitioning of the available resources with an increasing dominance of a few species. Therefore, the relationship observed here seems likely to be explained by the complementarity hypothesis.

Copyright © 2003 John Wiley & Sons, Ltd.     

Characterization of Amaranthus cruentus L. seed proteins by 2-DE and LC/MS–MS: Identification and cloning of a novel late embryogenesis-abundant protein

Amaranth is taking great attention as an important cereal crop that could fulfill food requirements for the growing population, especially in developing countries. However, the protein composition of these seeds is not well known yet. We have used the proteomics tools to characterize amaranth seed proteome. About 400 proteins spots were resolved on two-dimensional gel electrophoresis (2-DE) and protein spots were analyzed by LC-MS/MS (liquid chromatography tandem mass spectrometry). Identified proteins were related to stress and defense responses, metabolic, respiratory, and oxide-reduction processes. One abundant spot was identified as a Late Embryogenesis Abundant (LEA) protein and the gene was cloned and characterized. The AcLEA cDNA contains a 418 bp open reading frame (ORF) encoding a polypeptide of 139 amino acids. Bioinformatics analysis showed that AcLEA belongs to LEAs Group 5. Proteomics is a powerful technique that could be used even in non-sequenced organisms such as amaranth. The obtained information reveals that amaranth seed, beyond the classical seed storage proteins, contains proteins related to protection against stress. The identification of these proteins opens the door to the application of new strategies to improve the quality of amaranth production.     

Plant growth and survival of five introduced and two native/naturalized perennial grass genotypes exposed to two defoliation managements in arid Argentina

The field performance of the native Pappophorum vaginatum, the naturalized Eragrostis curvula and various cultivars of the introduced Achnatherum hymenoides and Leymus cinereus was evaluated as potential forage resources in rangelands of arid Argentina during the warm seasons of 2007/2008 and 2008/2009. Plants of these grass species, obtained from seeds, were transplanted to the field in 2006, when they were 1 year old. During the study years, there were two defoliation managements: plants of all study genotypes either remained undefoliated (controls) or were defoliated twice a year during spring at 5 cm stubble height. Despite tiller number being lower (P < 0·05) on defoliated than on undefoliated plants, and total leaf length per unit basal area being similar (P > 0·05) between defoliation managements by mid‐spring, there were no differences (P > 0·05) in dry weight production between defoliated and undefoliated plants in all genotypes at the end of the second growing season. Plants of one or more of the introduced genotypes showed a similar (P > 0·05) or greater (P < 0·05), but not lower, tiller number per plant and per square centimetre, daughter tiller production, total leaf length and dry weight production per unit basal area than the native species at the end of the first and/or second growing seasons. These morphological variables were similar (P > 0·05) or greater (P < 0·05) in the native than in the naturalized genotype. Plant survival, however, was lower (P < 0·05, overall mean = 20%) in the introduced than in the native (>70%) or naturalized (>39%) genotypes at the end of the first or second growing seasons.     

Tanjungides A and B: New Antitumoral Bromoindole Derived Compounds from Diazona cf formosa. Isolation and Total Synthesis

Tanjungides A (1) (Z isomer) and B (2) (E isomer), two novel dibrominated indole enamides, have been isolated from the tunicate Diazona cf formosa. Their structures were determined by spectroscopic methods including HRMS, and extensive 1D and 2D NMR. The stereochemistry of the cyclised cystine present in both compounds was determined by Marfey’s analysis after chemical degradation and hydrolysis. We also report the first total synthesis of these compounds using methyl 1H-indole-3-carboxylate as starting material and a linear sequence of 11 chemical steps. Tanjungides A and B exhibit significant cytotoxicity against human tumor cell lines.     

Biosynthetic Modularity Rules in the Bisintercalator Family of Antitumor Compounds

Diverse actinomycetes produce a family of structurally and biosynthetically related non-ribosomal peptide compounds which belong to the chromodepsipeptide family. These compounds act as bisintercalators into the DNA helix. They give rise to antitumor, antiparasitic, antibacterial and antiviral bioactivities. These compounds show a high degree of conserved modularity (chromophores, number and type of amino acids). This modularity and their high sequence similarities at the genetic level imply a common biosynthetic origin for these pathways. Here, we describe insights about rules governing this modular biosynthesis, taking advantage of the fact that nowadays five of these gene clusters have been made public (thiocoraline, triostin, SW-163 and echinomycin/quinomycin). This modularity has potential application for designing and producing novel genetic engineered derivatives, as well as for developing new chemical synthesis strategies. These would facilitate their clinical development.     

Developing scoring functions for soil quality to assess land suitability for irrigated wheat in Southern Algeria

Efficient land-use management could be optimized through appropriate advice regarding land suitability and achieving a reasonable yield while preserving soil and environmental conditions. The main objective of this study was to determine and define those locations capable of wheat cultivation with irrigation in the municipality of N’goussa (Southern Algeria). Expert opinion (EO) method was used for selecting minimum soil data set (MDS_-EO), where six influencing soil properties (sand, pH, salinity, organic matter, gypsum and lime) and other three key parameters (slope, land use/land cover, and proximity to roads and electricity network) were considered important for irrigated wheat cultivation in the study area. Indicators were weighted according to principal component analysis (PCA) loadings and summed. PCA revealed that the selected nine indicators are appropriate for accounting 73.63% of the total variance. Two maps of land suitability were created here (expressed as a soil quality index within a range from 0 to 1) from multivariate analysis and already known standard scoring functions (SSFs) by using two data sets (pure and scored). The distribution of soil quality index (IQI) showed that above 70% of the land was moderately (MS) to highly suitable (HS) for irrigated wheat. The estimation accuracy of soil quality classes has been evaluated using random forests (RF), having as a dependent variable the final class of land suitability used in each map. It was determined that estimation accuracy could reach 60% (pure data set) and 65% (scored data set) at 0.5 cut-off value for soil quality classes. Final prioritizing according to ranks of sensitivity index (SI) and efficiency ratio (ER) showed that IQI obtained from scored data set is the most suitable approach in soil quality assessment of arid areas. Therefore, it could be applied in southern Algeria for land management, integrated planning and environmental assessments.